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A variety of beneficial pharmacological activities have been reported for Se-enriched Grifola frondosa polysaccharides. However, little has been reported on its absorption, and its intestinal uptake and transport profiles remain unknown. Based on our previous research, the aim of this study was to investigate its absorption from two aspects - the polysaccharides and selenium of Se-enriched Grifola frondosa polysaccharides (Se-GFP-22) across Caco-2 cells in vitro. The Caco-2 cells monolayer culture model was successfully constructed to study the transport and uptake of Se-GFP-22. The results revealed that the uptake and transport of Se-GFP-22 were time- and concentration- dependent. Transport studies illustrated that Se-GFP-22 could penetrate Caco-2 cells, mainly mediated through the same routes as endocytosis and selenium in the organic selenium (Se-GFP-22) was more easily absorbed than that in the inorganic selenium control group (sodium selenite). The uptake of Se-GFP-22 may be a macropinocytosis pathway, which was an accumulation from cytoplasm to nucleus process. Se-GFP-22 was a moderately absorbed biological macromolecule testified by the apparent permeability coefficients (Papp) value and transport rates. This work illustrates the characteristics on uptake and transport of Se-GFP-22 and all these results may help to explore the mechanism of polysaccharide absorption in vitro.

Abelmoschus esculentus L (okra) is widely used as a healthy vegetable and favourable source of dietary medicine. Okra flowers which are by-products of okra, are rich in polysaccharide, polyphenols and trace elements etc., however, except a few for health tea, most of them were discarded as the waste of resources. In this study, a polysaccharide named AEFP22 was extracted, purified and identified from okra flowers, and its physicochemical property and antioxidant activity were also elucidated. AEFP22, with a molecular weight of 2.741 × 105 Da, was composed of Rha, GalA and Gal in the ratio of 1: 1.02: 0.86. The methylation and nuclear magnetic resonance (NMR) analysis indicated AEFP22 was composed of [2)-α-D-Rhap-(1 → 4)-α-D-GalpA-(1 → 2,4)-α-D-Rhap-(1 → 4)-α-D-GalpA-(1] with branch of terminal T-α-D-Galp pointed at C4 of 1,2,4-α-D-Rhap. The Conge-red test, Atomic force microscope (AFM) and Scanning electron microscope (SEM) further revealed the triple-helical conformation, irregular sheet structure with molecule aggregations of AEFP22. The physicochemical property analysis indicated AEFP22 possessed stable thermal property and exhibited shear-thinning and normal Newtonian fluid in different concentrations, −7.04 mV zeta potential and polymerization phenomenon existed in AEFP22 solution. AEFP22 exhibited good 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability. These results indicated potential utilization of AEFP22 in nutritional food and material application.

In this study, a Se-deficient mice model was successfully developed by feeding a Se-deficient diet (0.02 mg Se/kg diet) for 4 weeks, and Se supplementation by Se-polysaccharides (Se-GFP-22) was lasted for 4 weeks. The immunomodulatory activity and Se supplementation of Se-GFP-22 from Se-enriched G. frondosa was investigated. Results showed that Se-GFP-22 remarkably enhanced glutathione peroxidase (GSH-Px) and thioredoxin reductase (TrxR) activities in liver, kidney and plasma, and serum, liver, spleen and kidney Se levels of Se-deficient mice. Se-GFP-22 increased the thymus and spleen indices, phagocytic index, co-mitogenic (ConA- or LPS-stimulated) activities on splenocytes and DTH reaction. Se-GFP-22 caused significant increments in cytokine (IL-1β, TNF-α and IFN-γ) levels and Ig G, Ig A, Ig M and Ig E levels. Se-GFP-22 exhibited superior immunomodulatory effects than GFP-22. These findings indicated that Se-GFP-22 promote the protective effects against Se deficiency-induced immunosuppression and could be a potential immunomodulatory agent and a dietary Se-supplement.

The structural characterization, antioxidant activity and protective effect against H2O2-induced injury RAW264.7 cells of a new polysaccharide (MP1) isolated from Lepidium meyenii Walp. were investigated. The molecular weight was estimated to be 4.67 × 105 Da by HPLC-ELSD analysis. Monosaccharide composition was determined to be arabinose and galactose in a molar ratio of 2:1 by GC-MS and PMP-HPLC-UV analysis. Methylation combined with 1D and 2D NMR spectrum analysis revealed that MP1 was an arabinogalactan. MP1 possessed a moderate antioxidant activity in vitro in DPPH, ABTS, superoxide and hydroxyl radicals scavenging, Fe2+ chelating, lipid peroxidation inhibition and reducing power assays. MP1 could also effectively protect RAW264.7 cells from H2O2-induced injury by maintaining stable cell viability, decreasing ROS, MDA and LDH level and enhancing SOD, GSH-Px and CAT activity. These results suggested the potential utilization of MP1 as a natural antioxidant for food and pharmaceutical applications.

An efficient ultrasonic-cellulase-assisted extraction (UCE) of Cistanche tubulosa polysaccharide (CTP) was established. The response surface methodology based on Box-Behnken Design was employed to further optimize extraction conditions. After quaternary ammonium salt precipitation, the polysaccharide of C. tubulosa was characterized by different techniques. The results showed that a maximum polysaccharide yield of 22.31±0.45% was achieved at a pH of 5.2 for 31.5min at 54.1°C. Compared to hot water extraction, the yield of CTP in UCE and polysaccharide content increased to 44.96% and 70.13±2.19%, respectively. There was no marked difference among polysaccharides extracted using different methods from the infrared spectrum. Ultrasonic-cellulase-assisted extraction polysaccharide showed a fibrous structure from scanning electron microscopy and was composed of rhamnose, mannose, glucose, and galactose in a molar ratio of 2.18:1:28.29:1.43 by gas chromatography. The circular dichroism results indicated that polysaccharides had a maximum positive peak around 210nm with different peak values. The thermogravimetric analysis and differential scanning calorimetry were used to test the thermostability of CTP. Besides, CTP demonstrated appreciable antioxidant potential on antioxidant experiments in vitro. The results suggested that UCE is an effective method for CTP extraction and its polysaccharide showed appreciable antioxidant activity.

In our previous study, a polysaccharide was extracted from Schisandra Chinensis (Trucz.) Baill and found with anti-diabetic effects. The aim of this study was to investigate the anti-diabetic effects of the low weight molecular polysaccharide (SCPP11) purified from crude Schisandra polysaccharide and illustrate the underlying mechanism in buffalo rat liver cells. The insulin resistance model of BRL cells was established by incubating with insulin solution for 24h. The effects of SCPP11 on regulating related protein and mRNA expression in an insulin and AMPK signal pathway were investigated by western blot and RT-PCR analysis. SCPP11 showed no cytotoxicity to BRL cells and could improve the glucose consumption in BRL cells. SCPP11 increased the protein expression of Akt, p-AMPK and GLUT-4 in BRL cells. Moreover, SCPP11 could enhance the mRNA expression levels of IRS-1, PI3K, Akt, GLUT-4, AMPKα and PPAR-γ in BRL cells at the same time. In conclusion, SCPP11 possessed effects in improving glucose consumption by up-regulating the expression of GLUT-4 which might occur via insulin and AMPK signal pathway and could be a potential functional food to prevent and mitigate the insulin resistance condition.

A polysaccharide termed Se-GP11 was extracted and purified from Se-enriched Grifola frondosa in our previous study. This study investigated the characterization, anti-tumor and immunomodulatory activity of Se-GP11. The results showed that Se-GP11 was composed of mannose, glucose and galactose with a molar ratio of 1:4.91:2.41. The weight-average molecular weight (Mw) and weight-average mean square radius (Rw) of Se-GP11 in 0.1M sodium chloride solution were 3.3×10(4)Da and 32.8 nm. Se-GP11 existed as a globular conformation with random coil structure. Se-GP11 had no anti-tumor activity against HepG-2 cells in vitro, and it significantly inhibited the growth of Heps tumor in vivo. Se-GP11 increased the relatively thymus and spleen weights as well as serum necrosis factor-alpha (TNF-α) and interleukin-2 (IL-2) levels. In addition, Se-GP11 promoted the phagocytosis and NO production of RAW264.7 as compared with that of the normal control group. The results revealed that the Se-GP11 may exhibit the anti-tumor through improving immunologic function of the tumor bearing mice.

GP11 had been reported to have effectively anti-tumor activity by improving the immune function in our previous study. To avoid drawbacks of the 5-Fu, GP11 in combination with 5-Fu was investigated in this study. The results demonstrated that such synergism displayed enhance the anti-tumor activity of 5-Fu. Additionally, a strength effect was also observed in regulating immune function of GP11 and 5-Fu simultaneous administration, such as enhancing serum interleukin-2 (IL-2) and tumor necrosis factor-alpha (TNF-α) secretion, and increasing immune organs weights. Moreover, GP11 could improve the haematological and biochemical parameters deterioration, superoxide dismutase (SOD) activities reduction and malondialdehyde (MDA) levels enhancement in non-immune organs induced by 5-Fu. All these results illustrated that GP11 exhibited attenuated and synergized effect on 5-Fu by improving the immune function. It could be developed as an auxiliary preparation for chemotherapeutic drugs.

The structural characterization, antioxidant activity and protective effect against H

A novel bioactive polysaccharide, GFP-22, was isolated from the fruit bodies of Grifola frondosa by anion-exchange and gel filtration chromatography. Structure of GFP-22 was investigated using gas chromatography–mass spectrometry (GC–MS), methylation, nuclear magnetic resonance (NMR), high-performance size exclusion chromatography-multi-angle laser light scattering-refractive index detector (HPSEC-MALLS-RI) and atomic force microscopy (AFM) analysis. The backbone of GFP-22 is composed of 1,4-β- d -Glcp, 1,3-β- d -Glcp, 1,6-α- d -Glcp, 1,6-α- d -Galp, 1,4,6-α- d -Manp and 1,3,6-α- d -Manp units. Molecular weight of GFP-22 is 2.72 × 104 Da. GFP-22 has a linear filamentous structure. The administration of GFP-22 could improve or reverse the CTX-induced immunosuppression, significantly enhance the spleen and thymus indices, spleen lymphocyte proliferation and cytokines production in splenocytes. These findings suggest that GFP-22 could be explored as a natural and potential immunomodulatory agent and as an alternative means of lessening chemotherapy-induced immunosuppression.

Previous study revealed that Se-GP11 could exhibited its antitumor activity by improving the immune functions. 5-Fu, as a chemotherapeutic drugs, can kill many immune cells in addition to tumor cells. Accordingly, the enhanced antitumor and reduced toxicity of Se-GP11 on 5-Fu were estimated in this study. The results demonstrated that Se-GP11 could evidently increase the antitumor activity of 5-Fu. Furthermore, Se-GP11 could enhance the immune functions during the tumor inhibition process of 5-Fu for increasing the cytokines secretion (IL-2 and TNF-α) and immune organs weights. In addition, Se-GP11 could reduce the toxicity of 5-Fu on liver by improving the hematological and biochemical parameters and up-regulating the SOD activities and down-regulating the MDA levels. Taken together, the results indicated that Se-GP11 may develop as an auxiliary preparation to chemical antitumor drugs.

The chemical carboxymethylated polysaccharide (CMAAP), which is derived from the poorly water-soluble, crude polysaccharide from Auricularia auricula (AAP), was prepared. The degree of substitution of CMAAP was 0.857, and its solubility in water reached 0.6 mg/mL, much higher than that of AAP (0.1 mg/mL). One fraction, CMAAP22, was purified. Chemical analysis indicated that CMAAP22 was composed of mannose and glucose in a ratio of 1.06:1 with a molecular weight of 3.4 × 10(6)Da. IR spectra and (13)C NMR spectra indicated that the C2, C4 and C6 positions were partially substituted by -CH(2)COOH. AFM revealed that CMAAP22 was spherical particle structure with a diameter of 80-300 nm. Moreover, the antioxidant activity of CMAAP22 was improved, nearly twice as much as that of the AAP, especially in the scavenging of DPPH and ABTS+.

Schisandra chinensis (Turcz.) Baill has been used in traditional Chinese medicine for centuries. Previous studies have shown that Schisandra polysaccharide (SCPP11) has robust antitumor activity in vivo. In this study, the immunomodulatory activity and mechanisms of action of SCPP11 were investigated further to reveal its mechanism of action against tumors. Results showed that SCPP11 increased the thymus and spleen indices, pinocytic activity of peritoneal macrophages, and hemolysin formation in CTX-induced immunosuppressed mice. Moreover, SCPP11 significantly increased immunoglobulin levels, cytokines levels in vivo and induced RAW264.7 cells to secrete cytokines in vitro. RAW264.7 cells pretreated with SCPP11 significantly inhibited the proliferation of HepG-2 cells. In addition, SCPP11 promoted both the expression of iNOS protein and of iNOS and TNF-α mRNA. TLR-4 is a possible receptor for SCPP11-mediated macrophage activation. Therefore, the data suggest that SCPP11 exerted its antitumor activity by improving immune system functions through TLR-4-mediated up-regulation of NO and TNF-α.

Previous study indicated that the refined polysaccharide from Schisandra could improve the CTX-induced inhibition of T and B lymphocytes proliferation. Accordingly, the enhanced antitumor and reduced toxicity effects of a low molecular weight purified polysaccharide from Schisandra (SCPP11) were investigated in 5-fluorouracil (5-Fu) treated Heps-bearing mice. The results revealed that the SCPP11 (oral administration) exhibited a significant enhanced effect of antitumor activity when combined with 5-Fu. Moreover, a increased effect was also observed in boosting immunity functions when the Heps-bearing mice receiving SCPP11 combination with 5-Fu administration, including increased in thymus indexes and enhancing serum IL-2 and TNF-α secretion. In addition, SCPP11 could ameliorate the hematological and biochemical parameters changes induced by 5-Fu to normal level, and reduce the formation of MDA and enhance the activities of SOD in liver to against 5-Fu induced free radical damage. The above results suggested that the SCPP11 combined with 5-Fu presented enhanced effects on antitumor activity and the SCPP11 could attenuate the 5-Fu-induced toxicity effect. It could serve as a new and promising adjuvant for chemotherapy drugs.

In this study, hyaluronic acid-selenium (HA-Se) nanoparticles as novel complexes were synthesized and their antitumor activities in vivo were investigated. The mice inoculated with Heps tumor were orally administered with HA-Se nanoparticles at 86.45 mg/kg (H) and 4.32 mg/kg (L) body weights as high and low doses respectively (2.20% selenium content in the HA-Se nanoparticles samples by ICP-AES) for 10 days. The transmission electron microscopy (TEM) results indicated that the HA-Se nanoparticles were spherical with mean size of 50-70 nm. The HA-Se nanoparticles could significantly reduce tumor weights at the tumor inhibition ratios of 46.92% (H) and 49.12% (L) respectively. However, in the 5-fluorouracil positive group (25 mg/kg), the tumor inhibition ratio was 61.71%. From the study, the HA-Se nanoparticles (4.32 mg/kg) significantly increased thymus and spleen relative weights, enhanced the activities of superoxide dismutase (SOD), reduced the formation of malondialdehyde (MDA) and the activities of aspartate transaminase, alanine transaminase and crea in Heps tumor mice. The results of the study indicated that the HA-Se nanoparticles are potential antitumor candidate for cancer treatment.

Grifola frondosa has been widely consumed in China and other Asian countries. Recent studies on G. frondosa have focused on the activities of polysaccharides extracted by water, and the activities of polysaccharides extracted by enzymolysis have not been studied. In this work, the relationship between the physical properties and antioxidant activity of polysaccharides extracted from G. frondosa by boiling water and enzymolysis was studied. Five polysaccharide extracts from the fruit body of G. frondosa were prepared by different extracting methods including boiling water, single enzyme enzymolysis with three different single enzymes (cellulose, pectinase, and pancreatin), and combined enzyme enzymolysis (cellulose:pectinase:pancreatin; 2:2:1). Characteristics such as the viscosity, Mw, polysaccharide content, protein content, infrared spectra, and antioxidant activities of the extracts were evaluated. The highest antioxidant activity was exhibited by the extracts prepared by combined enzyme extraction. The correlation analysis between antioxidant activity and polysaccharide content, protein content, Mw or viscosity indicated that the Mw had a more important role in antioxidant activity. Overall, the results indicate that the combined enzyme polysaccharide extracts can be developed as a new potential natural antioxidant.